In 46 patients with type II diabetes mellitus (DM). the plasma pancreatic polypeptide (PP) response to intramuscular injection of 0.2μg/kg caerulein (Ce) was studied in special reference to the exocrine pancreatic function. As a result, the possibility was raised that pancreatic juice regulates PP secretion. Also, mongrel dogs were subjected to common bile duct drainage (BD group), pancreatic duct drainage (PD group), pancreatic duct and common bile duct drainage (PBD group), or sham operation (Control), and plasma PP responses to intravenous Ce infusion (10 ng/kg/min, 20 min) were investigated to ascertain the effect of pancreatic juice and bile upon PP secretion. Exocrine pancreatic function was examined using oral PFD and p-aminobenzoic acid (PABA) tolerance tests, then collecting 6-hr urine specimens and determining the recovery of PABA in urine, represented, as PFD/PABA ratio. For plasma PP responses, the amount of PP release was calculated by subtraction (A-B), where A was the area under the plasma PP response curve for 120 min after Ce loading (ng·min/ml) and B was the basal PP level (ng/ml)×120 (min).
The findings are, briefly as follows:
1) Plasma PP responses were significantly lower in DM patients with disturbed exocrine pancretic function (n=19) than in those with normal function (n=19), and there was a significant positive correlation between the amount of PP release and exocrine pancreatic function in all DM patients (p<0.02).
2) Multiple regression analysis, was performed to seek a possible relationship between the amount of PP release arid sex, age, mode of treatment, duration of DM, ophthalmofundoscopic findings, HbA1, 24-hr urinary excretion of CPR, PFD/PABA ratio, or the coefficient of variation of R-R interval in ECG. The analysis showed that only the PFD/PABA ratio had a significant influence on plasma PP response.
3) The amount of PP release by the PD or PBD group, which were treated to prevent the inflow of pancratic juice into the duodenum, was significantly lower than that of the sham operated or BD group respectively.
The above results indicate that the amount of PP release depends upon the output of pancreatic juice.

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In order to determine whether diabetes mellitus is a high risk factor for cholelithiasis, abdominal sonographic examination was performed to screen for cholelithiasis in 176 diabetics and 241 non-diabetic controls. The incidence of cholelithiasis was higher in diabetcis than in non-diabetics (17.6% vs. 7.5 %). Then gallbladder contractility was investigated to determine the possible mechanism of development of cholelithiasis in diabetics. The gallbladder contractile rate was estimated by using real-time ultrasonography to measure the changes in the size of the gallbladder after oral egg-yolk administration or intramuscular injection of caerulein. Sixty minutes after egg-yolk loading, the gallbladder contracted gradually by 57.0% and 55.9% respection in 10 non-diabetics and 9 diabetics without any signs of diabetic neuropathy. In 8 diabetics with neuropathy, the contractility was reduced (34.9% at 60 min). With caerulein injection in 9 non-diabetcis, the gallbladder contracted by 61.3% at 30 min and then dilated gradually. In 7 diabetics without neuropathy it contracted well but did not dilate thereafter. In 10 diabetics with neuropathy, it either contracted poorly or remained uncontracted.
Since patients with somatostatinoma are known to have both diabetes and cholelithiasis, the relationship between somatostatin concentration in plasma and gallbladder contraction was investigated in diabetics. With high doses (50 ng/min/kg) of somatostatin infusion, contraction of the gallbladder by egg-yolk or caerulein was completely inhibited in 10 healthy volanteers. With low doses (1 ng/min/kg) of somatostatin infusion, gallbladder contraction by caerulein injection was inhibited in three of five normal volanteers, even when the palsma levels of somatostatin remained at about 20-30 pg/ml. Moreover, the plasma somatostatin levels in a few diabetics were about 20-30 pg/ml.
In conclusion, the high incidence of cholelithiases in diabetics seems to be due to reduced gallbladder contraction which is related to diabetic neuropathy and/or to incrased plasma levels of somatostatin.

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A computer program is described giving recommendations concerning the initial strength (expressed in pulse rate) of exercise for diabetics. Major emphasis on safety has been incorporated in the recommendations. The knowledge data base was constructed concisely from medical publications. Then a computer program written in list was produced using a soft-ware (ESHELL), which helps users to adopt a suitable system. The data items should be clearly defined, since the computer program in general, does not allow for ambiguity. The definitions were made in order to produce safer computer recommendations.
Thirty six questions were constructed unnecessary questions were eliminated from the questionaire. After the execution of the program, a recommendation suited to the clinical state of the patient was produced. When the program was applied to twenty clinical cases, recommendations consistent with those from medical doctors were obtained. However, in eight out of the twenty cases, there were differences in the explantations for the recommendations. This may be because doctors made judgements about a patient from all angles, while in the computer program, the most important basis for the recommendation was the cardiovascular state of the subject. When this kind of computer program is applied in a clinical situation, it is necessary to recognize that the final responsibility lies with the physician who uses the soft ware and takes care of the patient. If more rules were added to this prototype program for the exercise therap for diabetics, the actual clinical judgement could be simulated. Then this kind of computer program would be a useful tool for diabetic clinics.

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The effects of elastase on lipid metabolism, platelet function, and blood coagulability were evaluated in 20 patients with diabetes mellitus. Blood was sampled before medication, and at 8 and 16 weeks after oral administration of 10800 units of elastase per day. Measurements were made of platelet count, platelet sensitivity to ADP-aggregation, serum concentrations of total cholesterol, HDL-cholesterol and triglyceride, and plasma concentrations of β-thromboglobulin (β-TG), fibrinogen (Fbg) and antithrombin III (AT III).
After the administration of elastase, HDL-cholesterol increased (0 W: 46.9±12.1 mg/dl; 8 W: 50.3±11.1 mg/dl, p<0.001; 16 W: 53.2±11.9 mg/dl, p<0.001), and triglyceride decreased (0W: 151.4±49.2 mg/dl; 8W: 128.1±44.1 mg/dl, p<0.05; 16 W: 125.4±43.4 mg/dl, p<0.05).There was no significant change in total cholesterol. The platelet count was elevated and β-TG decreased after the medication (0W: 117.0±67.1 ng/ml: 8W: 91.9+60.5 ng/ml, p<0.01; 16 W: 72.7±54.5 ng/ml, p<0.001); however, platelet sensitivity to ADP-aggregation remained unchanged. Although both Fbg and AT III increased, the degree of increase in AT III (0W: 25.8±4.0 mg/dl; 8 W: 31.1±5.3 mg/dl, p<0.001; 16 W: 32.0±4.2 mg/dl, p<0.001) was greater than that in Fbg (0W: 421.0±76.8 mg/dl; 8W: 471.3±95.9 mg/dl, p<0.01; 16W: 470.6±104.2 mg/dl, p<0.05).
In conclusion, elastase improved lipid metabolism, suppressed platelet release reaction and increased the AT III level. Therefore, it appears that elastase is useful drug for the protection of vascular complications in diabetes mellitus.

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We studied the screening method for glucose tolerance without glucose drinking and with the use of a combination of fasting glucose (FPG) and hemoglobin A_1c (HbA_1c), which have recently been analyzed as routine tests in Japan. FPG analysis was limited to the enzymatic method and HbA_1c was analyzed by four methods with two different HbA_1c analyzers based on the principle of high performance liquid chromatography (HPLC). Although the accuracy and precision of glucose determination were guaranteed by the Expert Committee on Quality Control in JAHTS (Chairman: H. Kiyose), the interlaboratory differences in HbA1c determination were remarkable. HbA_1c levels were then recalculated by the SDI method using each average and standard deviation (X, SD). These Xs and SDs were calculated from the HbA_1c levels of examinees judged as normal by the 75 g oral glucose tolerance test (O-GTT) based on the recommendation of the Japan Diabetes Association (JDA). We specified the screening regions as follows: 1) FPG≥140 mg/dl, 2) HbA_1c≥X+2 SD, 3) FPG≥110 mg/dl-<140 mg/dl and HbA_1c≥X+1 SD. These conditions had to be satisfied. By using these criteria, the screening ratio was calculated as 17.1%, sensitivity 38.2%, specificity 42.2%, false positive 2.6% and false negative 25.5%. However, as the screening ratio was obtained from selected subjects, this ratio had to be corrected. Finally, the screening ratio was recalculated as 12.7% by the population of 75g O-GTT in 11, 569 randomized subjects.

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Binding and biological activities of a photoreactive insulin derivative [N^εB29-(2-nitro-4-azidophenylglycyl)-insulin] were characterized.
In the absence of UV-irradiation, the relative binding ability of the photoreactive insulin to insulin receptors was one-fourth that of native porcine insulin in U 937 monocyte-like cells (0.967 vs. 3.71%). The biological activity of the photoreactive insulin measured by 2-deoxy-glucose transport was correlated with the binding ability in rat adipocytes. By UV-irradiation the photoreactive insulin was crosslinked to its receptors and the structure of the insulin receptor of rat liver membrane was analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis and autoradiography. When gels were run under reduced conditions, the photoreactive insulin specifically labeled a protein of Mr. 135, 000.
The cross-linked photoreactive insulin to its receptor of rat liver membrane was incubated with the supernatant of IM-9 lymphocytes containing IDE, and the degradation was then measured by the TCA method. Activity of this supernatant to degrade the insulin-receptor complex was remarkably decreased compared with native porcine and photoreactive insulin (20.9, 47.7 and 48.0%, respectively, for two hours' incubation). These results suggest that this derivative appeared to be useful to study the post-binding process.

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The acid saline extract (ASE) of rat submaxillary gland powerfully degrades ¹²⁵I-glucagon. Besides ¹²⁵I-glucagon, ¹²⁵I-pancreatic polypeptide (PP) was also destroyed by ASE according to ordinary immunoassay system with trichloroacetic acid (TCA); but ¹²⁵I-insulin was intact in the presence of ASE. Leupeptin and, to a lesser extent p-chloromercuriphenylsulfonic acid (PCMS) and N-ethylmaleimide, inhibited the destruction of ¹²⁵I-glucagon or ¹²⁵I-PP according to TCA method. These facts were also confirmed by gel filtration of the assay mixture. In the presence of leupeptin (0.4 mM) and PCMS (16 mM), there was no shift of the peak of labelled glucagon or PP. Thus ASE degrades not only ¹²⁵I-glucagon but also ¹²⁵I-PP, and the thiol proteinase inhibitor has a strong inhibitory effect on them.

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A tetai of 858 patients (484 males and 374 females), who were admitted for diabetic education at Osaka Prefectural Hospital between 1970 and 1975, were followed up until the end of 1982, and their long-term prognosis and causes of death were studied.
1) Living patients at the end of observation numbered 582 (67.8%), the deceased accounting for 185 (21.6%).
2) The observed numbers of deaths were compared with the expected numbers calculated on the basis of sex-and age-specific mortality rates among the general population of Osaka. The Obs./Exp. ratios of all causes and renal disease were significantly increased in both sexes. The ratios of liver and pancreatic cancer and liver cirrhosis were significantly increased in males. On the other hand, a significantly increased ratio was found for female cardiovascular disease.
3) Next, a case-control study was conducted within this cohort, in order to elucidate risk factors related to deaths from liver cancer and cirrhosis. The case group was 28 diabetic patients with newly developed disease who died of liver cancer or cirrhosis. The control group was selected from living diabetic patients matched for sex, year of admission (±one year) and birth year (±5 years), on the basis of a case to control ratio of 1: 3 or 1: 2.
4) Serum transaminase activities were elevated in 85.7%(24/28) of cases and 29.3%(22/75) of controls. The relative risk was estimated to be 14.5 among diabetic patients with elevated transaminase activities.
5) Alcohol consumption was compared between the two groups. A strong positive association between drinking habits and deaths from liver cancer or cirrhosis was observed, and there was a significant dose-response relationship after adjustment for serum transaminase levels.
6) Neither hepatitis B virus nor blood transfusion was a major risk factor for deaths from liver cancer or cirrhosis among diabetic patiets. Rates of diabetic triopathy were comparable between cases and controls, and oral hypoglycemic agents did not seem to increase the risk of liver cancer or cirrhosis.
The above-mentioned findings may suggest that hepatitis B surface antigen negative hepatitis and alcohol consumption were major independent risk factors related to liver cancer and cirrhosis among diabetic patietns in Japan.

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In order to investigate the mechanism of insulin resistance, lymphocytes from various patients were transformed by Epstein-Barr virus. These cells were able to bind insulin. Decreased insulin binding was shown in three different types of cells from a 6-year-old boy with insulin resistance (JRM 1-a): erythrocytes (37.4% of normal), transformed lymphocytes (9.8% of normal) and fibroblasts (53.3% of normal). In addition to lymphocytes from JRM 1-a, those from patients with non-insulin dependent diabetes mellitus (NIDDM) and a patient with Type B insulin receptor disease were transformed. Although insulin binding was decreased in the erythrocytes from these subjects, insulin binding to the transformed lymphocytes was not altered. For further evaluation of the defect of insulin binding in the patient JRM1-a, two subunits of insulin receptor on transformed lymphocytes were studied. First, the alpha-subunit of insulin receptor was labeled with ¹²⁵I-insulin by a chemical cross linker. Second, the beta-subunit of lectin-purified insulin receptor was labeled with ³²P-ATP in the absence or presence of insulin (autophosphorylation). Both alpha-subunits and beta-subunits of insulin receptor from JRM 1-a showed normal molecular size on the autoradiographs of sodium dodecyl sulfate poly acrylamide gels, but their contents of labeled materials were decreased compared with those of the controls. But insulin (500 ng/ml) stimulated ³²P-ATP incorporation into the beta-subunit of transformed lymphocytes from JRM 1-a to the same extent as in the control (630% and 550% over basal, respectively). In spite of decreased insulin binding to fibroblasts from JRM 1-a, the maximally insulin-stimulated glucose incorporation into his fibroblasts was not altered. However, the insulin dose-response curve of JRM 1-a cells was shifted to the right. These results indicated that the defects resided in insulin bindng but not in the post-receptor steps in the patient JRM 1-a, and that investigation of transformed lymphocytes and fibroblasts is useful for the clarification of insulin receptor abnormalities.

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The assay of furosine, an acid-hydrolysis product derived from fructose-lysine in glycated protein, was used as a specific method for determining Amadori compounds using high-performance liquid chromatography.
The nenenzymatic glycation of hair protein, which is readily taken non-invasively from the living body, was used as an indicator of blood glucose control.
We investigated the correlation between the furosine levels in hair of 1-cm lengths from the scalp and the fasting blood glucose level, as well as the correlation between the former and hemoglobin A1c (HbA1c) levels at the time of hair sampling.
The HbA1c level was well correlated with the furosine level in hair 0.5-1.5 cm, 1.5-2.5 cm and 2.5-3.5 cm from the scalp, but the corelation was best for the 0.5-1.5 cm hair portion. Hair in that section reflects blood glucose control during the preceding 1-1.5 months depending on the rate of hair growth.
The highly significant correlation between the furosine level in hair 0.5-1.5 cm from the scalp and the HbA_1c level suggests that the condition of blood glucose control at previous arbitrary time points can be estimated by using hair samples at various lengths from the hair root.

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A potent aldose reductase inhibitor, ONO-2235 (Ono Pharmaceuticals, Osaka, Japan) helps prevent peripheral nerve dysfunction in diabetes mellitus. To determine whether this compound is effective against diabetic renal damage, we compared urinary albumin excretion measured by radioimmunoassay and urinary N-acetyl-β-D-glucosaminidase (NAG) activity before and after treatment with ONO-2235. Urinary albumin excretion and NAG activity were measured twice with an interval of 5 months in diabetic patients without clinical proteinuria as determined with a reagent strip test. Fifteen patients who had both high urinary albumin excretion and high NAG activity in the two assays were given 150 mg of ONO-2235 daily by mouth for 8 weeks. The urinary albumin level significantly decreased in 11 patiens whose level was 200 mg per gram of creatinine or less 8 weeks after the treatment started (74.4±57.3 vs. 19.5±16.9 mg/g of creatinine, mean±SD, p<0.02). Four weeks after the treatment ended, urinary albumin excretion was measured in 7 of these patients, and each showed an elevation in urinary albumin which had been decreased by the aldose reductase inhibitor. However urinary albumin in 4 patients with albuminuria of over 200 was not decreased 8 weeks after the treatment started.
There was no significant difference in urinary NAG before and after treatment (10.4±3.6 vs. 11.8±4.8 U/g of creatinine). Neither fasting blood glucose nor hemoglobin A_1c changed during the study.
These results suggest that impaired polyol metabolism may participate in the development of diabetic renal disease and that aldose redutase inhibitor may be effective against diabetic renal damage.

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Multiple low-dose of streptozotocin induces autoimmune diabetes in mice which is characterized by hyperglycemia with lymphocytic infiltration into the pancreatic islets. The influence of genetic background on the susceptibility to this experimental form of diabetes was studied by comparing the susceptibilities of various congenic resistant and recombinant strains with a B 10 background. On the basis of the results of the first experiment, animals were injected on five consecutive days with streptozotocin (40 mg/kg, i.p.). The mice were considered diabetic when non-fasting serum glucose levels were above 200 mg/dl.
In congenic resistant strains, mice from strains B 10. (H-2^b) and B 10. BR (H-2^k) showed high incidences of diabetes in response to streptozotocin treatment. However, mice from strains B 10. D 2 (H-2^d) and B 10. S (H-2^s) showed low incidences of diabetes. Therefore, it is suggested that the haplotypes b and k are high-susceptiblity alleles and that d and s are low-susceptibility alleles.
In congenic recombinant strains, mice with the same haplotype on the K, E, S and D loci in the H-2 complex showed different susceptibilities, indicating that the diabetic susceptibility genes are located outside the K, E, S and D loci.
Mice from strains with the high-responder haplotypes k and b on the A locus in the H-2 complex showed high incidences but mice with the low-responder alleles d and s showed low incidences. The results suggested that genes coding for susceptibility to diabetes are located in the A locus within the H-2 complex.

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